Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
J Bacteriol. 2012 Apr;194(7):1789-99. doi: 10.1128/JB.06827-11. Epub 2012 Jan 27.
The streptococcal promiscuous plasmid pMV158 (5,540 bp) replicates by the rolling-circle mechanism and can be mobilized among a wide number of Gram-positive and -negative bacteria. The plasmid region involved in its conjugative transfer includes the mobM gene, which encodes the MobM relaxase, and the cis-acting origin of transfer (oriT). MobM initiates transfer by cleavage of supercoiled pMV158 DNA at a specific dinucleotide within oriT. In the present work, we have performed a detailed transcriptional analysis to assess the role of MobM in the control of its own gene expression. By in vivo and in vitro approaches, we demonstrated that mobM transcription in Escherichia coli was mostly initiated from a promoter (Pmob2) different from the one (Pmob1) used in Lactococcus lactis. Whereas promoter Pmob1 was embedded within the oriT sequence, promoter Pmob2 was placed apart from but adjacent to oriT. Further, MobM was able to repress the expression of its own gene from both promoters. Given the promiscuity of pMV158, the organization of the mobM promoter region suggests a strategy of the plasmid to cope with different transcription machineries of the hosts it colonizes.
链球菌杂合质粒 pMV158(5540bp)通过滚环机制复制,可在许多革兰氏阳性和阴性细菌中转移。涉及质粒接合转移的区域包括 mobM 基因,该基因编码 MobM 解旋酶和顺式作用转移起点(oriT)。MobM 通过在 oriT 内的特定二核苷酸处切割超螺旋 pMV158 DNA 来启动转移。在本工作中,我们进行了详细的转录分析,以评估 MobM 在自身基因表达调控中的作用。通过体内和体外方法,我们证明了大肠杆菌中 mobM 的转录主要是从不同于乳球菌(Lactococcus lactis)中使用的启动子(Pmob1)的启动子(Pmob2)开始的。虽然启动子 Pmob1 嵌入 oriT 序列内,但启动子 Pmob2 位于 oriT 之外但相邻。此外,MobM 能够从两个启动子抑制自身基因的表达。鉴于 pMV158 的混杂性,mobM 启动子区域的组织表明了质粒应对其定植的宿主不同转录机制的策略。